| ObjectiveBased on the potent antioxidant effects of reduced glutathione (GSH), we investigated the putative protective role of GSH by the level of toll-like receptor 4(TLR4), heat shock protein 70 (HSP70), and that of tumor necrosis factor-α(TNF-a) in liver against sepsis-induced hepatic injury in rats.MethodsSepsis-induced hepatic injury was induced by cecal ligation and puncture (CLP) in male Sprague-Dawley rats. Rats were randomly divided into 3 groups (n=24), which were sham group, CLP group and GSH treatment group. Rats of GSH treatment group were intravenously administered with GSH (300mg/kg) after the operation and the rats of other two groups were treated with same dose saline. Rats of each group were sacrificed at the time point of Oh,2h,6h and 24h after the operation (n=6). The samples of blood and liver tissues were harvested at each time point. TLR4, HSP70 and TNF-a in liver tissues were examined by ELISA kits and histological alterations by HE staining. The serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), direct bilirubin (DBIL) and total bilirubin (TBIL) activities were determined by auto analyzer.Results1,The serum levels of TBIL, DBIL, ALT and AST in rats:Compared with sham group, the levels of TBIL, DBIL, ALT and AST in CLP group were highly increased at 6h and 24h after operation (the serum levels of TBIL, DBIL, ALT and AST at 6h after operation were 3.82±0.48μmol/L, 2.05±0.36μmol/L,59.3±18.1U/L,129.5±21.1U/L in CLP group vs. 2.45±0.29μmol/L,1.05±0.21μmol/L,27.5±4.2U/L,47.2±7.8U/L in sham group, P<0.05). Compared with CLP group, the levels of TBIL, DBIL, ALT and AST in GSH treatment group (6h) were dominantly decreased (the serum levels of TBIL, DBIL, ALT and AST at 6h after operation were 2.82+0.52μmol/L,1.48μ0.31μmol/L,39.0±13.8U/L,84.5±17.9U/L in GSH treatment group vs.3.82±0.48μmol/L,2.05±0.36μmol/L, 59.3±18.1U/L,129.5+21.1U/L in CLP group, P<0.05).2,The levels of TLR4 and TNF-a in liver tissues: Compared with sham group, the levels of TLR4 and TNF-αin liver tissues in CLP group were highly increased at 2h and got the peak at 6h after operation (the levels of TLR4 and TNF-a at 6h after operation were 22.14±3.04ng/mL, 98.84±13.57pg/mL in CLP group vs.7.40±0.65ng/mL,12.65±1.96pg/mL in sham group, P<0.05). After GSH intervention, the levels of TLR4 and TNF-a in liver tissues were highly decreased at 6h after operation compared with CLP group (the levels of TLR4 and TNF-a at 6h after operation were 15.23±2.06ng/mL,54.24±9.53pg/mL in GSH treatment group vs.22.14±3.04ng/mL,98.84±13.57pg/mL in CLP group, P<0.05).3,The levels of HSP70 in liver tissues:Compared with sham group, the levels of HSP70 in liver tissues in CLP group were mildly increased at 2h,6h and 24h after operation (44.17±5.72ng/mL,51.71±7.03ng/mL, 36.45±7.13ng/mLinCLPgroupvs.17.09±2. 11ng/mL,16.36±2.39ng/mL, 16.33±1.84ng/mL in sham group, P<0.05). After GSH intervention, the levels of HSP70 in liver tissues were highly increased at 2h,6h and 24h after operation compared with CLP group (68.37±8.21ng/mL, 82.52±10.61ng/mL,77.05±14.85ng/mL in GSH treatment group vs. 44.17±5.72ng/mL,51.71±7.03ng/mL,36.45±7.13ng/mL in CLP group, P <0.05).4,Histopathological injuries were observed significantly in liver tissues in CLP group at 6h and 24h, characterized by inflammatory cells infiltration, profound cellular degeneration and severe vascular congestion. The histopathological changes of liver tissues in GSH treatment group were much milder than that of the CLP group.ConclusionThese data revealed that GSH could exert protective effects on sepsis-induced hepatic injury. And these beneficial effects appeared partly due to decrease TLR4 levels and increase HSP70 levels in liver tissues and thereby attenuating the production of TNF-a in liver tissues. Thus, supplementing antiseptic hepatic injury treatment with GSH may be beneficial in the clinical setting. |
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